The origin of the brown frogs with 2n=24 chromosomes

Late replication and C-banding analyses of somatic metaphase chromosomes were attempted on three species of brown frogs with 2n=26 chromosomes (Rana japonica, R. tsushimensis andR. temporaria), and three with 2n=24 chromosomes (R. ornativentris, R. dybowskii andR. chensinensis), which are distributed in the Palearctic region. The late replication banding patterns were highly conserved in these species. Four chromosome inversions were demonstrated inR. ornativentris, two inR. dybowskii and two inR. tsushimensis. From a detailed comparison of late replication and C-banding patterns between the 2n=26 and the 2n=24 species, it was found that an end-to-end fusion of two small chromosomes (nos 11 and 13) in an ancestral 2n=26 species may have produced the medium-sized no. 6 chromosome of the 2n=24 species.     

Rana esculenta complex: An experimental analysis of lethality and hybridogenesis

Summary Experiments designed to analyze the lethality and hybridogenesis in the European green frog complex have yielded the following results: 1. As a rule the inter-se cross ofRana esculenta is lethal, but several crosses have produced fully viable progeny. The frequency of such break-through crosses appears to be related to parental population structure. 2. Parabiotic joining of lethal to viable embryos indicates that manifestation of the lethal effect is autonomous. There is, however, a 16–18% increase in the life span of the lethal partner. 3. Studies of LDH isozyme patterns revealed that thelessonae-specific alleles coding for the B^a and B^c subunits can be passed to the F₁ progeny from a parental female or male of theesculenta phenotype. This demonstrates that there is no total elimination of thelessonae genome in theesculenta germ cells. 4. Immunologically, offspring from the inter-se cross ofR. esculenta show a closer relationship to theridibunda than to theesculenta phenotype. Variations of antigenic protein patterns suggest the possibility of chromosomal recombination betweenlessonae andridibunda in theesculenta hybrid. These results are confirmed by two-dimensional electrophoretic analysis of proteins in the oocytes of the three frog phenotypes.This work was supported by grants from the Swiss National Science Foundation and the Georges and Antoine Claraz Schenkung.     

Experimental gynogenesis provides evidence of hybridogenetic reproduction in theRana esculenta complex

Summary The gynogenetic offspring of the hybrid frogRana esculenta (R. ridibunda x R. lessonae) are exclusively of theridibunda type. This is due to the premeiotic exclusion of thelessonae genome from the hybrid's germ cells.Acknowledgments. Prof. M. Fischberg, Genève, and Prof. P. Vogel, Lausanne, provided laboratory facilities. This research was supported by the Swiss National Science Foundation, grants 3.221-0.73 and 3.644-0.75.     

Cytogenetic studies of Hynobiidae (Urodela). XIV. Analysis of the chromosome of a Chinese salamander, Batrachuperus pinchonii (David)

The chromosome number of a Chinese salamander, Batrachuperus pinchonii, was re-examined. Adults and embryonic specimens had a diploid number of 66, with 33 bivalents during meiosis, in contrast to previous reported results. Furthermore, when C-banding analysis was performed with embryos, chromosomes with banding patterns homoeologous to those of Salamandrella keyserlingii and Hynobius species were found. It appears, therefore, that Batrachuperus, Salamandrella and Hynobius might be derived from a common ancestral species in eastern Asia. Received 22 August 1997; received after revision 14 October 1997; accepted 20 November 1997     

Effets de l'actinomycine D sur le développement normal et sur les modifications expérimentales de la morphogénèse de l'oeuf de l'oursinParacentrotus lividus

Summary In the presence of actinomycin D (20–40 µg/ml), the development of the eggs of the sea urchin,Paracentrotus lividus, is slowed from the late morula and stopped at the blastula stage. The development is immediately stopped in the blastula treated with actinomycin D (20–40 µg/ml). The inhibitory effects of actinomycin D are prevented by deoxyribonucleic acid. Actinomycin D does not exert animalizing or vegetalizing effects. However, the enhancing of vegetalizing action of lithium and the weakening of animalizing effects of zinc ions and Evans blue have been observed in the presence of actinomycin D. These observations may reflect some difference in the state of dependence of differentiation of entomesodermic and ectodermic structures towards the nucleus.     

G-banding patterns of the housefly,Musca domestica,autosomes and sex chromosomes

Summary G-banding patterns were found on the X-chromosomes of the housefly. This indicates the presence of major gene loci on these chromosomes, i.e. sex-linked genes. G-banding patterns were also found on the Y-chromosome of 2 preparations. The first polyploid cell (a tetraploid, 2n=24) forMusca domestica was observed in one of the banding preparations.     

Plasma albumin patterns of the speciesRana ridibunda in Greece

Summary Serum electrophoretic studies on the speciesRana ridibunda displayed one new albumin band D and a total of 6 albumin patterns B, BC, C, BD, CD, BCD. The albumin patterns found are compared with those reported from Central and Northern Europe.Acknowledgments. Thanks are due to Prof. M. Kattoulas, Drs C. Triantaphillidis, H. E. J. Wijnands and Th. Sofianidou for assistance and/or valuable advice.     

Biochemistry of frog ribonucleases

Frogs have unique pyrimidine base-specific RNases, with structures similar to those of turtle, iguana and chicken RNases. Among the four frog RNases discussed here, three from Rana pipiens, R. catesbeiana and R. japonica oocyte cells show anti-tumour activity, and the latter two show lectin activity towards sialic acid-rich glycoproteins. In this review, (i) we compare their unique primary structures with respect to the locations of their disulphide bridges, three-dimensional structure, base specificity and heat stability as compared with RNase A, and (ii) we summarize current knowledge about the mode of action of lectin and the antitumour activities of the three frog RNases.     

Biochemical variation in theRana esculenta complex: A new hybrid form related toRana perezi andRana ridibunda

Summary Investigations of the green frogs from western Europe for electrophoretic variations at 4 enzyme loci demonstrated a new form which must be considered as a hybrid betweenRana ridibunda andR. perezi. Biochemical evidence supports the hypothesis that its reproduction is hybridogenetic, as it is forR. esculenta.     

Genetic control of LDH isozymes in theRana esculenta complex

Summary Studies of LDH isozymes in the European green frogs showed that the synthesis of the B subunits is controlled by 3 alleles at a single genetic locus. The genetic evidence supports the hypothesis thatRana esculenta is the hybrid ofR. lessonae x R. ridibunda.This work was supported by grants from the Schweizerischer Nationalfonds, the Georges und Antoine Claraz-Schenkung and the Karl Hescheler-Stiftung.     

Chromatin ofh regions of human chromosomes at high resolution

Summary Segmentation of the secondary constriction region (h) of human chromosomes 1, 9 and 16 is demonstrated by a high resolution banding technique. Based on these staining properties, it is suggested that the composition of theh region in human chromosomes is heterochromatic as well as euchromatic.     

C-banding pattern on the chromosomes of the Japanese house shrew,Suncus murinus riukiuanus,and its implication

Summary The C-band on the chromosomes of the Japanese house shrew,Suncus murinus riukiuanus (Insectivora), was studied. Various types of C-banding pattern were found in the genome of this subspecies. Such banding patterns could be useful for an understanding of autosome and sex-chromosome polymorphisms withinS. murinus.Acknowledgment. The authors are very grateful to Prof. T. Wasano of Fukuoka University for his encouragement, Dr K. Ogawa of Fukuoka University, the Nohara family for help in collecting specimens, and Prof. E.W. Jameson, Jr, University of California, Dr J.W. Bickham and his students of Texas A & M University for comments.     

The fate of intramitochondrial paracrystalline inclusion bodies in germ line cells of water frogs (Amphibia,Anura)

Summary Numerous intramitochondrial pararystalline inclusion bodies (ICIB) were observed in the germinal plasm of a mid-blastula, and in primordial germ cells (PGCs) after their migration to the germinal ridges, inRana ridibunda, R. lessonae andR. esculenta. In oogonia the number of ICIB decreases rapidly. Single ICIB are observed in the germ cells up to the leptotene stage; they have never been observed in pachytene oocytes. In diplotene oocytes that have reached a diameter of about 100 m ICIB are visible again, and their number increases concomitantly with oocyte growth.     

Vereinfachte Chromosomenbänderungsmethode bei landwirtschaftlichen Nutztieren

Zusammensetzung des Borax-Puffers Stammlösung A: Borsäure H₃BO₃MG 61,83, 0,2M=12,3 g/l 50 Teile Stammlösung B: Natriumborat NaBO₂–4 H₂O, MG 137,86 0,2M=27,57 g/l 50 TeileSusammenselzung des Sörensen-Puffers Sörensen-Puffer (nachSchnedl für Bandfärbung M/15 KH₂PO₄/Na₂HPO₄ mit einem pH von 6,8). KH₂PO₄ MG=136,09–1/15M=20,41 g/l Na₂HPO₄·2H₂O MG=177,99–1/15M=26,69 g/l.Der Gehalt an Methylen Violet Bernthsen in Giemsalösung ist von Bedeutung und kann durch leichtes Aufkochen aktiviert werden.

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Simple chromosome banding technique for farm animal investigations

Summary For routine cytogenetic investigations, a simple banding technique on chromosomes is described. Used on different cell material, avisible banding appearance is produced which makesthe typing of chromosomes precise and fast. On 2 examples of cattle and rabbit chromosomes, the identification of a centric fusion and a trisomic is shown.

     

Cytogenetic studies of Hynobiidae (Urodela). IX. Karyological characteristics ofHynobius abei Sato by means of R-and C-banding

Summary Chromosomal characteristics of the salamander speciesHynobius abei, from Ohimya (Kyoto) were revealed by the techniques of R-and C-banding. The karyotype ofH. abei was characterized by the shortness of an R-negative (C-positive) band in the terminal region of the long arm of chromosome 2 and a band encompassing the whole short arm of chromosome 10. These two bands inH. abei were the shortest among those of the variousHynobius species that have been examined. Otherwise no differences could be detected betweenH. abei and seven other pond-type species ofHynobius (2n=56) in terms of the banding patterns of 18 specifically identifiable pairs of their chromosomes.     

Isoforms of soluble alpha-tubulin in oocytes and brain of the frog (genus Rana): changes during oocyte maturation

Rana oocytes have previously been shown to contain much more soluble tubulin than does the brain, suggesting different assembly and disassembly dynamics of frog oocyte tubulin compared to that in brain. By using centrifugation, SDS-PAGE, two-dimensional gel electrophoresis and Western blots, probed with anti-α-tubulin monoclonal antibodies, polymorphic α-tubulins (isoforms) were compared in brains and follicle-enclosed oocytes of northern (Rana pipiens) and southern (R. berlandieri) frogs. Oocyte tubulin in both species had isoforms with greater ranges of isoelectric point (pI) than those of brain tubulins; in particular, the oocyte tubulin pIs ranged further into the acidic region of the isoelectric-focusing gels than corresponding brain tubulin. This difference may, in part, be responsible for the previously reported assembly differences between oocyte tubulin (undetectable assembly) and brain tubulin (high assembly). Isoforms of α-tubulin with relat ively acidic pI were more abundant in northern frog brain and oocyte soluble extracts than in analogous extracts from southern frogs. Furthermore, additional acidic α-tubulin isoforms were found in progesterone-treated oocytes (i.e., eggs), indicating increased heterogeneity of acidic a-tubulin isoforms during oocyte meiotic maturation. Among northern frog oocyte soluble components fractionated on Superose-6b columns, tubulin complexes with apparent molecular mass of about 1800 kDa were found to contain acidic α-tubulin isoforms while the putative oligomeric tubulins with an apparent molecular mass of about 250 kDa contained an additional relatively basic α-tubulin isoform. The acidic α-tubulin isoforms, therefore, are proposed to be associated with cold-adaptable cells of brain and oocytes, and may also be involved in stabilization of large soluble tubulin complexes in oocytes of the northern frog. Received 1 October 2002; accepted 9 October 2002 RID="*" ID="*"Corresponding author.     

Differenzierendes Kaulquappen-Fressen durch Molche

Summary Differential predation on tadpoles by newts (Triturus). 4 EuropeanTriturus species (T. alpestris, T. cristatus, T. helveticus, T. vulgaris) feed upon the tadpoles of the following Anura species which occur in sympatry with the newts:Rana esculenta, R. temporaria andBombina variegata. OnlyTriturus cristatus prey also upon the tadpoles ofBufo bufo andB. calamita. Triturus alpestris, T. helveticus andT. vulgaris do not eat anyBufo tadpoles. Even larvae ofTriturus alpestris prey upon tadpoles ofRana esculenta, R. ridibunda andBombina variegata but avoid tadpoles ofBufo calamita.

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Mit Unterstützung des Schweizerischen Nationalfonds zur Förderung der wissenschaftlichen Forschung.     

Hydrogen peroxide and hydroxyl radical involvement in the activation of caspase-3 in chemically induced apoptosis of HL-60 cells

Apoptosis of HL-60 cells induced by actinomycin D, H7, or daunorubicin was shown to involve the activation of caspase-3-like protease, 2 h after the addition of these drugs, based on microassay of enzyme activity by high-performance liquid chromatography. Catalase and a spin trap, N-t-butyl--phenylnitrone, which effectively inhibited the apoptosis induced by these drugs, also inhibited the activation of caspase-3-like protease. These results suggest that hydrogen peroxide and the hydroxyl radical are common mediators of caspase-3 activation caused by these chemicals, with apparently different functional mechanisms. Based on mitochondrial activity determined by oxygen consumption, complexes I, II, and IV were inhibited by actinomycin D. H7 inhibited complexes I and IV, 1 and 1.5 h respectively, after the addition of the drug to HL-60 cells. Daunorubicin inhibited complex IV, 1.5 h after the addition of the drug to HL-60 cells. Inhibition of complex IV by actinomycin D, H7, and daunorubicin were almost fully restored by the addition of cytochrome c. The release to the cytosol of cytochrome c by these drugs was also demonstrated by Western blot analysis. Addition of catalase inhibited the depression of complex IV activity induced by actinomycin D and H7. These observations indicate a direct relationship between hydrogen peroxide and the release of cytochrome c during apoptosis caused by actinomycin D, H7, and daunorubicin. Received 24 November 2000; received after revision 2 January 2001; accepted 30 January 2001